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Method for analyzing periodontal ligament stem cell osteoblastic differentiation by application of gene chip

A technology for the differentiation of periodontal ligament stem cells and osteoblasts, which is applied in biochemical equipment and methods, and the determination/inspection of microorganisms, and can solve the problem of inability to efficiently promote the differentiation of stem cells and bone lineages.

Inactive Publication Date: 2015-04-01
福建医科大学附属口腔医院
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Simply controlling one molecule or even one pathway will not be able to effectively promote the osteogenic differentiation of stem cells

Method used

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  • Method for analyzing periodontal ligament stem cell osteoblastic differentiation by application of gene chip
  • Method for analyzing periodontal ligament stem cell osteoblastic differentiation by application of gene chip

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Experimental program
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Effect test

Embodiment 1

[0019] Methods of using gene chip to analyze the differentiation of periodontal ligament stem cells into osteoblasts:

[0020] (1) Culturing periodontal ligament stem cells: Take out the periodontal ligament from the molars removed due to orthodontics and impaction under aseptic conditions, then cut the periodontal ligament into small pieces, and use type I collagenase and dispase enzyme in 37 Digested at ℃ for 1 hour, then centrifuged and resuspended, filtered the resuspension with a 100μm steel mesh to obtain a single cell suspension;

[0021] (2) Sorting of periodontal ligament stem cells: Use trypsin digestion to collect the periodontal ligament cells in the logarithmic growth phase of the single cell suspension obtained in step (1), and use STRO-1-PE according to the antibody specification Label the cells, collect STRO-1 positive cells by FACS flow cytometry, and the collected STRO-1 positive cells are the required periodontal ligament stem cells;

[0022] (3) Directional diffe...

Embodiment 2

[0025] The specific application of gene chip analysis of osteoblast differentiation miRNA expression profile:

[0026] experiment material

[0027] Among the selected people, the premolars extracted due to orthodontics were used clinically between the ages of 19 and 29 due to orthodontic and impacted molars, and the periodontal ligament was removed under aseptic conditions, cut, and then obtained. Periodontal ligament tissue is used as cell culture.

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Abstract

The invention provides a method for analyzing periodontal ligament stem cell osteoblastic differentiation by the application of a gene chip. A periodontal ligament stem cell is prepared to obtain a single-cell suspension, a periodontal ligament cell in logarithmic phase is sorted from the single-cell suspension; and by a STRO-1-PE marker, STRO-1 positive cells are sorted and collected through a FACS flow cytometer so as to obtain the required periodontal ligament stem cell. Then, the collected periodontal ligament stem cell undergoes clone culture, and when the periodontal ligament stem cell grows to 80% collection degree, pancreatin therein is also digested to obtain a seed solution. The seed solution is inoculated in a 6-well plate or a 24-well plate. After 24 hours, an induced liquid is used for culture. Periodontic treatment analysis by a gene chip is of profound significance for detecting changes of miRNA expression profile during the PDLSC osteoblastic differentiation process.

Description

【Technical Field】 [0001] The invention relates to a method for analyzing the differentiation of periodontal ligament stem cells into osteoblasts, in particular to a method for analyzing the differentiation of periodontal ligament stem cells into osteoblasts by applying a gene chip. 【Background technique】 [0002] According to the statistics of the American Association for Dental and Maxillofacial Research, 80% of the elderly over 70 suffer from periodontitis. However, periodontitis is a common cause of periodontal tissue defects and eventually tooth loss. Not only the decline in dental function, but also a serious impact on people's health and quality of life. Therefore, how to use tissue engineering methods to ensure periodontal tissue regeneration has become a hot topic for domestic and foreign scholars. [0003] The periodontal ligament stem cell (PDLSC) is the most direct and reliable regeneration seed cell in periodontal tissue. It is not only the most basic and important lin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6837C12Q1/6851C12Q2525/207C12Q2531/113C12Q2545/101
Inventor 吕红兵郑碧琼雷丽珊张明
Owner 福建医科大学附属口腔医院
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